Spearow J L, Trost B A
Department of Meat and Animal Science, University of Wisconsin, Madison 53706.
Biol Reprod. 1987 Oct;37(3):595-605. doi: 10.1095/biolreprod37.3.595.
This manuscript reports the development of a rapid, sensitive, and specific-enzyme linked immunosorbent assay (ELISA) suitable for measuring luteinizing hormone (LH) in cattle, sheep, rats, and mice. The LH ELISA used #15 anti-LH serum-coated 96-well plates and peroxidase-labeled bovine luteinizing hormone (bLH). Bovine LH labeled with peroxidase by the periodate method was stored at -15 degrees C for over 20 mo without appreciable loss of activity. With bLH-B5 used as the standard diluted in assay buffer, the LH ELISA had a sensitivity of 79.6 +/- 31 pg/ml, and a 50% displacement point of 359 +/- 69 pg/ml. With rat LH-RP-2 (rLH) used as the standard diluted in assay buffer, the LH ELISA had a sensitivity of 102 pg/ml and a 50% displacement point of 531 pg/ml. The LH ELISA was highly specific for LH from several mammalian species. This LH ELISA was seven times more sensitive for bLH than a radioimmunoassay (RIA) with comparable sample volumes. The LH ELISA was validated for measurement of LH in buffer, tissue culture media, and sera. Depending on the sensitivity desired, the LH ELISA can be conducted in 3 to 48 h, produces no hazardous waste, and can easily be automated. Use of this LH ELISA offers improvements in speed, convenience, economy, sensitivity, and safety over comparable RIA procedures. The LH ELISA was also conducted with other monoclonal and polyclonal antibodies to LH. The LH ELISA can be performed with other LH antisera, provided the antisera has a high affinity and specificity for LH and can be uniformly coated on 96-well plates.
本手稿报告了一种快速、灵敏且特异的酶联免疫吸附测定法（ELISA）的开发，该方法适用于测定牛、羊、大鼠和小鼠体内的促黄体生成素（LH）。LH ELISA使用#15抗LH血清包被的96孔板和过氧化物酶标记的牛促黄体生成素（bLH）。通过高碘酸盐法用辣根过氧化物酶标记的牛LH在-15℃下储存超过20个月，活性无明显损失。以在测定缓冲液中稀释的bLH-B5作为标准品，LH ELISA的灵敏度为79.6±31 pg/ml，50%置换点为359±69 pg/ml。以在测定缓冲液中稀释的大鼠LH-RP-2（rLH）作为标准品，LH ELISA的灵敏度为102 pg/ml，50%置换点为531 pg/ml。该LH ELISA对几种哺乳动物物种的LH具有高度特异性。与具有可比样品体积的放射免疫测定法（RIA）相比，这种LH ELISA对bLH的灵敏度高7倍。该LH ELISA已被验证可用于测定缓冲液、组织培养基和血清中的LH。根据所需的灵敏度，LH ELISA可在3至48小时内完成，不产生有害废物且易于自动化。与可比的RIA程序相比，使用这种LH ELISA在速度、便利性、经济性、灵敏度和安全性方面都有改进。还使用了其他针对LH的单克隆和多克隆抗体进行LH ELISA。只要抗血清对LH具有高亲和力和特异性并且可以均匀地包被在96孔板上，就可以用其他LH抗血清进行LH ELISA。
